An integrated approach for sediment characterization of Dead Sea, Israel: Implications to marine environment.

The Dead Sea (DS) sediment samples from two different sites on the west coast were analyzed. The objective is to document sediment characterization in order to study the interrelation existing among ingredients. The analyses show that Fe, Mg and Zn are the most significant trace elements found in the Sediment. Anthropogenic activities such as Industries, untreated sewage, agricultural wastes are some of the pollution sources that are considered to be lethal to the marine ecosystem. The DS is slowly disappearing at an alarming rate of 1 m/year. Hence a constant and precise mitigation, monitoring and management is mandatory. The present work is part of such goal in order to understand the health of the DS ecosystem. A cumulative and consistent effort of the Governments, stakeholders, tourists and the public will help in order to preserve and protect the health of the rare DS marine ecosystem for future generations.

Converse magneto-electric effects in a core-shell multiferroic nanofiber by electric field tuning of ferromagnetic resonance.

This report is on studies directed at the nature of magneto-electric (ME) coupling by ferromagnetic resonance (FMR) under an electric field in a coaxial nanofiber of nickel ferrite (NFO) and lead zirconate titanate (PZT). Fibers with ferrite cores and PZT shells were prepared by electrospinning. The core-shell structure of annealed fibers was confirmed by electron- and scanning probe microscopy. For studies on converse ME effects, i.e., the magnetic response of the fibers to an applied electric field, FMR measurements were done on a single fiber with a near-field scanning microwave microscope (NSMM) at 5-10 GHz by obtaining profiles of both amplitude and phase of the complex scattering parameter S11 as a function of bias magnetic field. The strength of the voltage-ME coupling Av was determined from the shift in the resonance field Hr for bias voltage of V = 0-7 V applied to the fiber. The coefficient Av for the NFO core/PZT shell structure was estimated to be - 1.92 kA/Vm (- 24 Oe/V). A model was developed for the converse ME effects in the fibers and the theoretical estimates are in good agreement with the data.

Benthic foraminifera as potential ecological proxies for environmental monitoring in coastal regions: A study on the Beypore estuary, Southwest coast of India.

The benthic foraminiferal diversity index was computed from Beypore estuary sediments. The abundance and diversity of Quinqueloculina lata, Textularia agglutinans, Haplophagmoides canariensis, and Quinqueloculina stelligera were dominated by stress-tolerant taxa such as Ammonia tepida, A.parkinsoniana, Nonion grateloupi, and N. scaphum in the estuary. The small-size foraminifera probably perished in a juvenile stage because of the high temperature and low salinity that prevailed in the ecosystem. The dominance of stress tolerant benthic foraminifera and absence of Elphidium species in the estuary suggest the prevalence of hypoxic (low-oxygen) conditions. The consistent low-diversity index of foraminifera indicates that the ecosystem is moderate to highly stressed ecologically in the Beypore estuary. The application of benthic foraminifera as a bioindicator for assessing the environmental stress in the Beypore estuary is key in monitoring these fragile coastal ecosystems.

Data on Molluscan Shells in parts of Nellore Coast, southeast coast of India.

X-ray diffraction (XRD), Scanning Electron Microscope-Energy Dispersive Spectroscopy (SEM-EDS), and Fourier Transform Infrared Spectroscopy (FT-IR), were applied to analyze the organic matrix of two Molluscan shells. The Mollusca shells are mineral structure and calcium carbonate crystallized as aragonite. The FT-IR spectra showed Alkyl Halide, Alkanes, Alcohols, Amides, Aromatic, and Hydroxyl groups in the organic matrix of the whole (organic and mineral) Molluscan shells. SEM images of particles of the two Molluscan shells at different magnifications were taken. The morphologies of the samples show a flake like structures with irregular grains, their sizes are at micrometric scale and the chemical analysis of EDS indicated that the major elements of Cardita and Gastropoda were C, O, and Ca, consistent with the results of XRD analysis. The results of the analysis of the EDS spectra of the shells showed that the content of most of the powder composition of shells is the element carbon, calcium oxygen, aluminium, and lead peaks that appear on the Cardita and Gastropoda and shells powders tap EDS spectra. The present work examined organic matrix of the selected shells of the heavily polluted and light polluted sites, along Nellore Coast, South East Coast of India. The heavily polluted sites have significantly thickened shells. The data demonstrated the sensitivity of this abundant and widely distributed intertidal fragile environment.

Dynamics of Pulicat Lake mouth analysis using geospatial data, east coast of India: Implications to socio-economic scenarios.

Pulicat Lake is one of the major wetlands in India. It is the second largest brackish water lagoon in India next to Chilika Lake in Orissa state. Pulicat Lake sits beside the Bay of Bengal so, the study on the mouth is vital. The investigations were carried out by using multi-temporal satellite imageries of IRS P6, LISS III data for four years viz., 2009, 2011, 2012 and 2013. Subsequent changes in the width of the lake at the southern side were measured. It is found that the lake mouth is not static but dynamic predominantly fluctuating year by year. Obviously, this poses threat to the lake biodiversity. Hence, it is high time to mitigate, manage, monitor and protect the existing width of the sea mouth to keep the lake biological, ecological, economically active. This paper noticed a considerable change in the mouth of the lake studied using satellite imageries and socio-economic settings.

Organic matter from benthic foraminifera (Ammonia beccarii) shells by FT-IR spectroscopy: A study on Tupilipalem, South east coast of India.

Fourier Transform Infrared Spectroscopy (FTIR) was used to study the variations in organic matters of benthic foraminifera (Ammonia beccarii) from four samples collected from beach environments from brackish environments along Tupilipalem coast (South east coast of India). Common absorption bands were observed as peaks in the range of 3600-3400 cm-1, 3000-2850 cm-1, 1750-1740 cm-1, 1640-1600 cm-1, 1450-1350 cm-1, 885-870 cm-1 and 725-675 cm-1 in all the shells of Ammonia beccarii. The FTIR spectrum of station-1 represents the presence of alkanes (CH3) and alkyl halide (C-F stretching) with absorptions at the range 1385-1255 and 1350-1150 cm-1 were observed and ether (C-O stretching) absorption band was observed at stations 1 and 3 with wavenumber of 1115 cm-1 and 1117 cm-1 respectively. Alkynes C-H bend was observed at station-1 with the wavenumber of 667.43 cm-1. The shifting of peak positions in all the samples is could be due to presence of organic matter in the samples. Satellite remote sensing and field observation data revealed that the river mouth at Tupilipalem coast was closed by a sand bar. Consequentially, this waterbody may affect the species diversity. •Positions of the sampling locations were identified using a hand-held Garmin Global Positioning System (GPS).•Foraminifera from the sediment were obtained using a mixture of Bromoform and Acetone.•The functional groups present in the benthic foraminifera shells were recorded in the spectral range of 4000-400 cm-1 using an FT-IR Spectrophotometer.

Heavy metal pollution monitoring with foraminifera in the estuaries of Nellore coast, East coast of India.

A total of 112 bottom water and sediment samples collected at fixed stations in pre-monsoon and post-monsoon from four estuaries (Pennar, Uppateru, Swarnamukhi, and Kalangi) showed foraminiferal test abnormalities in heavy metal concentrations (Co, Cr, Cu, Fe, Mn, Ni, and Pb). Low diversity of fauna was due to the predominance of a limited number of opportunistic species capable of achieving high densities in adverse environmental conditions and the reduction in the number of species intolerant of such conditions. In this study, classification of 54 common species according to their distribution is presented. Approximately 15 species showed quite low diversities at stations 23-27 and 44-51. Because of the effect of heavy metal pollution in these estuaries, drastic changes in the number of species and diversity of foraminifera were observed. These changes in foraminiferal species and the increase in test abnormalities are proxies of environmental stress on the estuarine ecosystem.

20ß-hydroxysteroid dehydrogenase gene promoter: potential role for cyclic AMP and xenobiotic responsive elements.

Teleostean 20ß-hydroxysteroid dehydrogenase (20ß-HSD) is involved in final oocyte maturation and steroid hormone metabolism. It has structural and functional similarities to mammalian carbonyl reductases that are involved in the metabolism of endogenous carbonyl and xenobiotic compounds. To understand the transcriptional regulation of 20ß-HSD, here we report the cloning of 20ß-HSD promoter from two fish species, rainbow trout and air-breathing catfish. Analysis of the promoter motifs, in silico identified the presence of several sites for transcription factor binding including cAMP, xenobiotic and steroid hormone responsive elements. Luciferase reporter assays with progressive deletion constructs demonstrated that 20ß-HSD type B of trout has no promoter activity while 20ß-HSD type A of trout and catfish 20ß-HSD promoters showed basal promoter activity. A TATA box flanked by a CAAT box is important for basal transcription. Deletion of cAMP responsive element in the promoter decreased basal promoter activity significantly. Reporter assays with forskolin and IBMX, drugs that increase intracellular cAMP induced the promoter activity over the basal level. Intriguingly, ß-nafthoflavone, an arylhydrocarbon receptor ligand, induced the 20ß-HSD promoter activity and is further evidenced by the induction of 20ß-HSD expression in the livers of catfish, in vivo. These results demonstrate for the first time that 20ß-HSD expression is not only modulated by cAMP but also by xenobiotics and further studies may provide significance to the ubiquitous distribution and broad substrate specificity of this enzyme.

Expression and immunolocalization of 20ß-hydroxysteroid dehydrogenase during testicular cycle and after hCG induction, in vivo in the catfish, Clarias gariepinus.

The maturation inducing hormone, 17α,20ß-dihydroxy-4-pregnen-3-one (17α,20ß-DP) is required for the meiotic maturation and is produced from the precursor 17α-hydroxyprogesterone by the enzyme 20ß-hydroxysteroid dehydrogenase (20ß-HSD) in several teleosts. Central role of 20ß-HSD in ovarian cycle and final oocyte maturation is well studied when compared to spermatogenesis. In the present study, we investigated the localization and expression of 20ß-HSD in testicular cycle and gonadotropin induced sperm maturation. During testicular ontogeny, 20ß-HSD expression was detectable at 50 and 100 days post-hatch (dph), while the expression was high at 150 dph. In testicular cycle, highest levels of mRNA and protein of 20ß-HSD were observed during spawning phase. Intraperitoneal injection of human chorionic gonadotropin (hCG) to prespawning catfish elevated both 20ß-HSD transcripts and protein levels when compared to saline treated controls in a time-dependent manner. Serum 17α,20ß-DP levels, measured during different phases of testicular cycle as well as following the treatment of hCG, showed a positive correlation with the expression of 20ß-HSD. Immunolocalization revealed the presence of 20ß-HSD protein predominantly in interstitial cells and spermatogonia/spermatocytes while 20ß-HSD was undetectable in haploid cells (spermatids/sperm). These results together with high expression during spawning phase of testicular cycle and after hCG treatment in the prespawning catfish suggests a pivotal role for 20ß-HSD during testicular recrudescence leading to sperm maturation. Further studies using various fish models on testicular 20ß-HSD may provide interesting details to understand its importance in teleostean spermatogenesis.

Electrical and magnetic properties of nanocrystalline BiFeO3 prepared by high energy ball milling and microwave sintering.

A new synthesis route with high energy ball milling and microwave sintering is used to obtain nanocrystalline BiFeO3 with improved dielectric and magnetic properties. Electrical and magnetic properties are compared with a conventionally sintered microcrystalline BiFeO3. It is found that the dielectric constant is increased more than one order of magnitude, electrical resistivity by six orders of magnitude and remnant polarization value is increased by 4-5 times for nanocrystalline BiFeO3 in comparison to conventionally sintered microcrystalline BiFeO3. Nanocrystalline BiFeO3 is seen to have ferromagnetic behavior whereas microcrystalline BiFeO3 is known to be antiferromagnetic.

A role for cytochrome P450 17alpha-hydroxylase/c17-20 lyase during shift in steroidogenesis occurring in ovarian follicles prior to oocyte maturation.

Cytochrome P450 17alpha-hydroxylase/c17-20 lyase (P450c17) is regarded as one of the key enzymes involved in the steroidogenic shift that occurs prior to oocyte maturation in teleosts. Role of P450c17 in the shift in steroidogenesis during oocyte maturation is a contentious issue even after identification of a novel type of P450c17 that lacks lyase activity. To understand the role of P450c17 in steroidogenic shift explicitly, a full length cDNA encoding p450c17 from ovary of air-breathing catfish, Clarias gariepinus was cloned. p450c17 transiently expressed in COS-7 cells converted progesterone to androstenedione through 17alpha-hydroxyprogesterone and catfish p450c17 was found to be expressed ubiquitously with relatively higher levels in gonads, brain, kidney and gills. Immunocytochemical analysis revealed the presence of P450C17 in follicular layer of ovarian follicle, interstitial cells and spermatocytes of testis. p450c17 expression and ratio of lyase to hydroxylase was high in preparatory and pre-spawning phases of ovary and low in spawning phase. Expression of p450c17 correlated well with testicular recrudescence with maximum expression in preparatory and spawning phases. Neither protein expression nor lyase/hydroxylase activity changed significantly during hCG-induced oocyte maturation, in vitro and in vivo though mRNA levels increased. These results tend to suggest that the ovarian follicles attains capacity to produce maximum precursor steroid levels before spawning that might contribute to the shift in steroidogenesis.

Cloning and expression of StAR during gonadal cycle and hCG-induced oocyte maturation of air-breathing catfish, Clarias gariepinus.

Complementary DNAs encoding steroidogenic acute regulatory protein (StAR) have been isolated from different fish species, yet the relevance of StAR during gonadal cycle and more importantly in final oocyte maturation has not been assessed so far. A cDNA encoding StAR was isolated from the ovarian follicles of air-breathing catfish, Clarias gariepinus. Catfish StAR exhibited 55 to 72% identity at nucleotide level with other vertebrate orthologs. RT-PCR analysis of tissue distribution pattern demonstrated the presence of StAR mRNA in various tissues including gonads, kidney, liver, brain and intestine of catfish. Real-time RT-PCR analysis revealed high expression of StAR mRNA in the pre-spawning phase of ovary while it was low in preparatory, spawning and regressed phases. In testis, maximum expression was noticed during the preparatory phase. During human chorionic gonadotropin (hCG)-induced oocyte maturation, both in vitro and in vivo, StAR mRNA levels were augmented by 2 h and then declined gradually to reach basal levels by 12 h as that of saline-treated controls. Taken together, high level of expression during hCG-induced oocyte maturation vis-à-vis in spawning suggests a role for StAR, in addition to the steroidogenic enzyme genes in final oocyte maturation.

New evidences for the involvement of 20beta-hydroxysteroid dehydrogenase in final oocyte maturation of air-breathing catfish.

20beta-hydroxysteroid dehydrogenase (20beta-HSD) synthesizes 17alpha,20beta-dihdroxy-4-pregnen-3-one, the steroid required for resumption of prophase-I arrested oocytes in teleosts. Though 20beta-HSD cDNAs have been cloned from few fish species, its role in final oocyte maturation (FOM) is still questionable. To study the role of 20beta-HSD in FOM more explicitly, we cloned and characterized 20beta-HSD from ovary of air-breathing catfish, Clarias gariepinus. Interestingly, Escherichia coli expressed recombinant proteins, both full-length and an N-terminal truncated proteins catalyzed the reduction of steroids and xenobiotics, however there was significant difference between them. Semi-quantitative RT-PCR and Western blots demonstrated the presence of 20beta-HSD transcript and protein in various tissues with relatively higher level in gonads, gill, kidney and brain. A positive correlation of 20beta-HSD expression was observed in different phases of ovarian cycles. Immunocytochemical/immunofluoroscence analysis with specific antibody identified presence of 20beta-HSD in follicular layer of ovary. Real-time RT-PCR and Western blotting showed an induction of 20beta-HSD expression during human chorionic gonadotropin (hCG)-induced oocyte maturation, in vitro and in vivo. Concomitantly, a rise in 20beta-HSD enzyme activity was also noticed. Specific inhibitors of carbonyl reductase inhibited not only recombinant protein catalytic activity but also hCG-induced oocyte maturation in a dose-dependent manner as evidenced by blocking of germinal vesicle break down. These results together provide new evidences for the involvement of 20beta-HSD in the FOM/meiotic maturation.

Novel 3beta-hydroxysteroid dehydrogenases from gonads of the Nile tilapia: phylogenetic significance and expression during reproductive cycle.

Multiple forms of 3beta-hydroxysteroid dehydrogenase/isomerase (3beta-HSD) and their differential tissue expression pattern have not been shown in any lower vertebrates. In the present study, we report cloning of two novel 3beta-HSDs and two variants from gonads of the Nile tilapia. 3beta-HSD cDNAs encode two peptides of 375 (3beta-HSD type-I/variant 1) and 367 (3beta-HSD type-II/variant 1) amino acid residues that share 31.9% homology. 3beta-HSD type-I/variant 1 shared high homology with other piscine counterparts while 3beta-HSD type-II/variant 1 exhibited homology to mammalian DeltaC27-3beta-HSD and multifunctional viral 3beta-HSD. The latter seems to be ancient form among vertebrates. Transiently transfected 3beta-HSDs' open reading frames in COS-7 cells converted exogenous pregnenolone/androsta-5-ene-3beta-17beta-diol to progesterone/testosterone. Tissue distribution pattern of 3beta-HSDs by RT-PCR revealed varied expression pattern. Northern blot analysis of 3beta-HSDs demonstrated steady or gradual rise in transcripts level at different gonadal stages. These data revealed the importance of novel 3beta-HSDs in teleosts and also provided phylogenetic significance.

Seabream GnRH immunoreactivity in brain and pituitary of XX and XY Nile tilapia, Oreochromis niloticus during early development.

Seabream gonadotropin-releasing hormone (sbGnRH)-the chief preoptic area-hypothalamus (POA-H) form of GnRH in tilapia is involved in sexual maturation. In this study, we investigated the qualitative changes in ontogeny of sbGnRH immunoreactivity (ir-), between sexes to understand its impending role during sex differentiation. For this, the differences in immunocytochemical localization of sbGnRH in genetically male (XY) and female (XX) fish were studied from 1 day after hatching (dah), through the critical period of sex differentiation (7-21 dah) to 40 dah and mature Nile tilapia. Specific antisera against sbGnRH were used for immunolocalization. SbGnRH ir- neurons were observed in POA-H as early as 5 and 15 dah in XY fish and XX fish, respectively. Higher ir- was detected in the POA-H of XY tilapia compared with XX population till 10 dah. There was a qualitative drop in sbGnRH ir- neurons/cell bodies in POA-H around 20 dah till 30 dah in XY population compared with other durations. SbGnRH ir- cells were detected in pituitary of XX fish by 15 dah and in XY fish around 10 dah but seemed to drop down by 20 dah in XY whereas it continued to remain steady in XX fish. The sbGnRH ir- in XY fish showed a rise from 35 dah and thence till 40 dah. This study revealed subtle differences in POA-H and pituitary sbGnRH ir- during early development between genetic male and female fish with possible implications in sex differentiation.

Spark plasma sintered Sm(2)Co(17)-FeCo nanocomposite permanent magnets synthesized by high energy ball milling.

Nanocomposite Sm(2)Co(17)-5 wt% FeCo magnets were synthesized by high energy ball milling followed by consolidation into bulk shape by the spark plasma sintering technique. The evolution of magnetic properties was systematically investigated in milled powders as well as in spark plasma sintered samples. A high energy product of 10.2 MGOe and the other magnetic properties of M(s) = 107 emu g(-1), M(r) = 59 emu g(-1), M(r)/M(s) = 0.55 and H(c) = 6.4 kOe were achieved in a 5 h milled and spark plasma sintered Sm(2)Co(17)-5 wt% FeCo nanocomposite magnet. The spark plasma sintering was carried out at 700 °C for 5 min with a pressure of 70 MPa. The nanocomposite showed a higher Curie temperature of 955 °C for the Sm(2)Co(17) phase in comparison to its bulk Curie temperature for the Sm(2)Co(17) phase (920 °C). This higher Curie temperature can improve the performance of the magnet at higher temperatures.

Thiourea-induced thyroid hormone depletion impairs testicular recrudescence in the air-breathing catfish, Clarias gariepinus.

We used thiourea-induced thyroid hormone depletion as a strategy to understand the influence of thyroid hormones on testicular recrudescence of the air-breathing catfish, Clarias gariepinus. Treatment with 0.03% thiourea via immersion for 21 days induced hypothyroidism (thyroid hormone depletion) as evidenced by significantly reduced serum T(3) levels. Thiourea-treated males had narrowed seminiferous lobules with fewer spermatozoa in testis, very little or no secretory fluid, reduced protein and sialic acid levels in seminal vesicles when compared to controls. The histological changes were accompanied by reduction in serum and tissue levels of testosterone (T) and 11-ketotestosterone (11-KT), a potent male specific androgen in fish. Qualitative changes in the localization of catfish gonadotropin-releasing hormone (cfGnRH) and luteinizing hormone (LH, heterologous system) revealed a reduction in the distribution of immunoreactive neuronal cells and fibers in thyroid depleted fish. Interestingly, thiourea-withdrawal group showed physiological and histological signs of recovery after 21 days such as reappearance of spermatozoa and partial restoration of 11-KT and T levels. These data demonstrate that thyroid hormones play a significant role in testicular function of catfish. The mechanism of action includes modulating sex steroids either directly or through the hypothalamo (GnRH)-hypophyseal (LH) axis.

Effect of methyl testosterone- and ethynyl estradiol-induced sex differentiation on catfish, Clarias gariepinus: expression profiles of DMRT1, Cytochrome P450aromatases and 3 beta-hydroxysteroid dehydrogenase.

The objective of the present study is to observe the effect of exogenous steroids, methyl testosterone (MT) and ethynyl estradiol (EEL) on gonadal differentiation and analyze its effect on the expression of several genes during testicular and ovarian differentiation in juvenile catfish. Exogenous hormone treatments (MT and EEL) were given by immersion at different days of hatching. The histological analysis revealed that the EEL- and MT-treatments resulted in the initiation of ovarian and testicular differentiation, respectively. This is further supported by specific expression of two forms of DMRT1 in the MT-treated group but not in the EEL-treated group at 47 days after hatching (dah). The reverse is true for the expression of ovarian aromatase. Results of the semi-quantitative RT-PCR show that brain aromatase transcript levels are high in 47 dah control (histologically female) and 47 dah EEL-treated fish, as compared to 47 dah MT-treated fish. At 60 dah, brain aromatase showed elevation in its expression. Interestingly, the expression pattern of 3 beta-HSD did not show any change in EEL- and MT-treated fish. The present study also provides a strategy to study sex differentiation, for those species where genetic sex population is unavailable.

Seabream GnRH: partial cDNA cloning, localization and stage-dependent expression in the ovary of snake head murrel, Channa striatus.

Vertebrate reproduction is under the neuroendocrine control of the hypothalamic decapeptide GnRH which synchronizes various reproductive events and influences other reproduction related aspects like spawning behavior and pheromonal action in fish. Multiple forms of GnRH peptides have been reported across diverse vertebrate and invertebrate classes. Here we report the partial seabream GnRH (sbGnRH) cDNA sequence cloned from the brain of Channa striatus (snake head murrel) a fresh water perciform with immense economic and medicinal value across Asiatic countries. sbGnRH mRNA was found in brain, gill and ovary of mature murrel with possible implications to the effect of GnRH on pheromonal phenomena and on reinitiation of oocyte meiosis. In keeping with the earlier reported role of GnRH in initiation of oocyte meiosis we here present evidence from RT-PCR, ICC demonstrating an increase in the level of sbGnRH mRNA in ovary from pre-vitellogenic to post-vitellogenic follicles.

Expression of 20beta-hydroxysteroid dehydrogenase and P450 17alpha-hydroxylase/c17-20 lyase during hCG-induced in vitro oocyte maturation in snake head murrel Channa striatus.

Partial cDNAs encoding carbonyl reductase like 20beta-hydroxysteroid dehydrogenase (20beta-HSD) and P450 17alpha-hydroxylase/c17-20 lyase (CYP17) were isolated from the ovary of snake head murrel and they exhibited high sequence identity to the Nile tilapia and rainbow trout, respectively. A low transcript level of both 20beta-HSD and CYP17 were detected in pre-vitellogenic follicles, while the transcript level was high in full-grown immature follicles. In hCG-induced in vitro oocyte maturation, we found a significant increase in 20beta-HSD transcript level after 2 h. The CYP17 transcripts also showed a considerable increase following hCG-induction compared to saline-treated controls. On the other hand, Western blot analysis demonstrated no significant change in the CYP17 protein level during hCG-induced in vitro oocyte maturation. Taken together, we suggest that in addition to 20beta-HSD, the CYP17 might have a role in the shift in steroidogenesis during meiotic maturation of snake head murrel.